Rat (Rattus) NGFB ELISA Kit

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Antigen
Nerve Growth Factor beta (NGFB) ELISA Kits
  • NGFB
  • ngfb
  • ns:zft386
  • xx:zft386
  • zgc:109730
  • hsan5
  • beta-ngf
  • Ngfb
  • Beta-NGF
  • HSAN5
  • beta-NGF
  • nerve growth factor (beta polypeptide)
  • beta-Nerve growth factor
  • nerve growth factor
  • NGF
  • ngf
  • FPV072
  • FPV076
  • Ngf
Reactivity
Rat (Rattus)
Alternatives
Kits with alternative reactivity to:
22
15
12
5
3
2
2
2
1
Method Type
Sandwich ELISA
Detection Range
15-15000 pg/mL
Minimum Detection Limit
15 pg/mL
Application
ELISA
Options
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Catalog No. ABIN625190
$ 476.30
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
11.217061 ABIN416255 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.6-1000 pg/mL 15.6 pg/mL Log in to see 5
11.217061 ABIN411337 Sandwich ELISA Cell Culture Supernatant, Serum 15.6-1000 pg/mL 15.6 pg/mL Log in to see 5
11.217061 ABIN1979400 Sandwich ELISA Cell Lysate, Tissue Lysate 15-15000 pg/mL 15 pg/mL Log in to see
11.217061 ABIN2708673 Sandwich ELISA Cell Culture Supernatant, Serum 15.6-1000 pg/mL 15.6 pg/mL Log in to see
11.217061 ABIN2113130 Cell Culture Supernatant, Serum Log in to see
1 ABIN3032862 Competition ELISA Cell Culture Supernatant, Plasma, Serum 20.6-15000 pg/mL 20.6 pg/mL Log in to see
1 ABIN579194 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN5594872 Sandwich ELISA Biological Fluids, Plasma, Serum, Tissue Homogenate 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN2685072 Sandwich ELISA Cell Culture Supernatant, Serum 15.6-1000 pg/mL 15.6 pg/mL Log in to see 5
1 ABIN5652206 Sandwich ELISA Biological Fluids, Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate 15.6 pg/mL - 1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN1112766 Sandwich ELISA 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN1847019 Serum, Plasma, Urine, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN5596649 Sandwich ELISA Log in to see
1 ABIN2945654 39.063-2500 pg/mL 39.063 pg/mL Log in to see
1 ABIN2116280 Sandwich ELISA 15.6-1000 pg/mL 15.6 pg/mL Log in to see

General

Antigen Nerve Growth Factor beta (NGFB) ELISA Kits
Reactivity Rat (Rattus)
Kits with alternative reactivity to:
(22), (15), (12), (5), (3), (2), (2), (2), (1)
Method Type Sandwich ELISA
Detection Range 15-15000 pg/mL
Minimum Detection Limit 15 pg/mL
Application ELISA
Pubmed 4 references available
Supplier Log in to see

Product Details NGFB ELISA Kit

Target details Application Details Handling References for NGFB Kit (ABIN625190) Images
Purpose Rat beta-NGF ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Detection Method Colorimetric
Analytical Method Quantitative
Specificity This ELISA kit shows no cross-reactivity with the following cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, TIMP-1, TNF-alpha.
Sensitivity < 15 pg/mL
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis

Target details

Product Details NGFB ELISA Kit Application Details Handling References for NGFB Kit (ABIN625190) Images back to top
Antigen
Alternative Name beta-NGF (NGFB ELISA Kit Abstract)
Background Brain-derived neurotrophic factor (BDNF) (Fragment)
UniProt Q06225
Pathways NF-kappaB Signaling, RTK Signaling, Regulation of Cell Size

Application Details

Product Details NGFB ELISA Kit Target details Handling References for NGFB Kit (ABIN625190) Images back to top
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation 1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
2. Sample dilution: Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of cell culture supernates. Suggested dilution for normal serum/plasma: 2 fold*. * Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
3. Assay Diluent B should be diluted 5-fold with deionized or distilled water before use.
4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 150 µL beta-NGF standard (50 ng/mL) from the vial of Item C, into a tube with 350 µL Assay Diluent A or 1x Assay Diluent B to prepare a 15,000 pg/mL standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 15,000 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 150 µL standard + 350 µL 200myl 200 µL 200 µL 200 µL 200 µL 15,000 5,000 1,666 555.6 185.2 61.73 20.58 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 500-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 500-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Assay Procedure 1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
5. Discard the solution. Repeat the wash as in step
6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7. Discard the solution. Repeat the wash as in step
8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Rat beta-NGF concentration (pg/mL) O D =4 50 (n m ) 0.01 0.1 1 10 10 100 1,000 10,000 100,000 Assay Diluent B Rat beta-NGF concentration (pg/mL) O D =4 50 (n m ) 0.001 0.01 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of beta-NGF is typically less than 15 pg/mL.
Recovery: Recovery was determined by spiking various levels of Rat beta-NGF into serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 79.32 69-89 Plasma 79.33 65-92 Cell culture media 107.7 94-117
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 106.5 102.6 102.6 Range ( %) 95-114 91-113 92-112 1:4 Average % of Expected 105.3 113.6 115.0 Range ( %) 94-114 102-123 104-126
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only

Handling

Product Details NGFB ELISA Kit Target details Application Details References for NGFB Kit (ABIN625190) Images back to top
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months

References for NGFB Kit (ABIN625190)

Product Details NGFB ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Tse, Novikov, Wiberg, Kingham: "Intrinsic mechanisms underlying the neurotrophic activity of adipose derived stem cells." in: Experimental cell research, Vol. 331, Issue 1, pp. 142-51, 2015

Faroni, Calabrese, Riva, Terenghi, Magnaghi: "Baclofen modulates the expression and release of neurotrophins in schwann-like adipose stem cells." in: Journal of molecular neuroscience : MN, Vol. 49, Issue 2, pp. 233-43, 2013

Faroni, Calabrese, Riva, Terenghi, Magnaghi: "Baclofen modulates the expression and release of neurotrophins in schwann-like adipose stem cells." in: Journal of molecular neuroscience : MN, Vol. 49, Issue 2, pp. 233-43, 2013

Reid, Sun, Wiberg, Downes, Terenghi, Kingham: "Nerve repair with adipose-derived stem cells protects dorsal root ganglia neurons from apoptosis." in: Neuroscience, 2011

Images

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Supplier Images
ELISA image for Nerve Growth Factor beta (NGFB) ELISA Kit (ABIN625190) Nerve Growth Factor beta (NGFB) ELISA Kit